[Histonet] Re: Histonet Digest, Vol 83, Issue 46
Feher, Stephen
sfeher <@t> CMC-NH.ORG
Mon Nov 1 10:51:27 CDT 2010
http://www.cytopathnet.org/tiki-index.php
Steve
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Miranda
D. Felton
Sent: Saturday, October 30, 2010 1:43 PM
To: histonet <@t> lists.utsouthwestern.edu
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Re: Histonet Digest, Vol 83, Issue 46
Does anyone know of a cytology listserv?
Thank you
Miranda
Sent from my iPod
On Oct 30, 2010, at 1:00 PM, histonet-request <@t> lists.utsouthwestern.edu
wrote:
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> Today's Topics:
>
> 1. RE: lymph nodes peeling out of OCT (Monfils, Paul)
> 2. PLP fixative (Truscott, Tom)
> 3. Precipitating Silver (Sheila Haas)
> 4. RE: Precipitating Silver (Monfils, Paul)
> 5. RE: PLP fixative (WILLIAM DESALVO)
> 6. IHC Staining Problem (Jennifer Hill)
> 7. Laura Miller is Out of the Office.
> (Laura.Miller <@t> leica-microsystems.com)
> 8. RE: RE: breast fixation times (Patsy Ruegg)
> 9. cryostat question (historsd <@t> aol.com) 10. Re: IHC Staining
> Problem (Jennifer MacDonald)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Fri, 29 Oct 2010 13:13:44 -0400
> From: "Monfils, Paul" <PMonfils <@t> Lifespan.org>
> Subject: RE: [Histonet] lymph nodes peeling out of OCT
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>
> <4EBFF65383B74D49995298C4976D1D5E075E08F9 <@t> LSRIEXCH1.lsmaster.lifespan.
> org
> >
>
> Content-Type: text/plain; charset="us-ascii"
>
> I place all tissues in liquid OCT for at least 15 minutes before
> freezing. This virtually eliminates the problem of tissue sections
> separating from the OCT after sectioning.
>
>
>
>
>
> ------------------------------
>
> Message: 2
> Date: Fri, 29 Oct 2010 10:55:45 -0700
> From: "Truscott, Tom" <ttruscot <@t> vetmed.wsu.edu>
> Subject: [Histonet] PLP fixative
> To: "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>
> <44F1D6D7EB8CC84F92859EE5C4E6ECB4011B20F1B562 <@t> CVMMBX.vetmed.wsu.edu>
> Content-Type: text/plain; charset="us-ascii"
>
> I am seeking basic information on PLP fixative. What is it? Can I make
> it or buy it? How long does fixation take for lymph node?
> Thanks in advance, Tom Truscott
>
>
> ------------------------------
>
> Message: 3
> Date: Fri, 29 Oct 2010 10:56:14 -0700 (PDT)
> From: Sheila Haas <micropathlabs <@t> yahoo.com>
> Subject: [Histonet] Precipitating Silver
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <450265.9811.qm <@t> web57802.mail.re3.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Has anyone heard of precipitating silver out of a solution by adding
> table salt?
> I read this somewhere as a means of disposing of silver easily. Any
> info would be appreciated.
> Thanks,
>
> Sheila Haas
> Laboratory Supervisor
> MicroPath Laboratories, Inc.
>
>
>
>
> ------------------------------
>
> Message: 4
> Date: Fri, 29 Oct 2010 14:02:26 -0400
> From: "Monfils, Paul" <PMonfils <@t> Lifespan.org>
> Subject: RE: [Histonet] Precipitating Silver
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>
> <4EBFF65383B74D49995298C4976D1D5E075E090C <@t> LSRIEXCH1.lsmaster.lifespan.
> org
> >
>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Yes, it is easy to do. Since silver chloride is insoluble in water,
> chloride added to a solution containing ionic silver will cause the
> white compound to precipitate out immediately. It will settle out
> onto the bottom of the container overnight, or can be filtered out.
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-
> bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sheila Haas
> Sent: Friday, October 29, 2010 1:56 PM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Precipitating Silver
>
> Has anyone heard of precipitating silver out of a solution by adding
> table salt?
> I read this somewhere as a means of disposing of silver easily. Any
> info would be appreciated.
> Thanks,
>
> Sheila Haas
> Laboratory Supervisor
> MicroPath Laboratories, Inc.
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 5
> Date: Fri, 29 Oct 2010 12:52:06 -0600
> From: WILLIAM DESALVO <wdesalvo.cac <@t> hotmail.com>
> Subject: RE: [Histonet] PLP fixative
> To: <ttruscot <@t> vetmed.wsu.edu>, histonet
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <BAY151-w591FEECED7C9459D94D71391450 <@t> phx.gbl>
> Content-Type: text/plain; charset="iso-8859-1"
>
>
> Follow the links for a receipe.
>
> www.pathology.ufl.edu/~molecular/PLP%20Fixative.doc
>
> http://www.niehs.nih.gov/research/atniehs/labs/lep/path-support/immuno
> /reagents.cfm
>
> www.hopkinsmedicine.org/.../multimedia/text_documents/Recipes_For_Maki
> ng_PLP_Fixative.doc
>
>
> William DeSalvo, B.S., HTL(ASCP)
>
>
>
>
>
>> From: ttruscot <@t> vetmed.wsu.edu
>> To: histonet <@t> lists.utsouthwestern.edu
>> Date: Fri, 29 Oct 2010 10:55:45 -0700
>> Subject: [Histonet] PLP fixative
>>
>> I am seeking basic information on PLP fixative. What is it? Can I
>> make it or buy it? How long does fixation take for lymph node?
>> Thanks in advance, Tom Truscott
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> ------------------------------
>
> Message: 6
> Date: Fri, 29 Oct 2010 20:05:13 +0000
> From: Jennifer Hill <jhill <@t> vet.k-state.edu>
> Subject: [Histonet] IHC Staining Problem
> To: histonet <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>
> <8AA2173DC209CA438077A832FF98BD7F02806442 <@t> VETMXHT.ads.vet.k-state.edu
> >
> Content-Type: text/plain; charset="iso-8859-1"
>
> I hope someone can help me with a problem I've been having with my
> CD79a hand stain. This is a stain that was validated and working
> perfectly, when a few months ago I started having reduced, to no
> staining on my controls, with the occasional run staining a little
> stronger. I followed the discussion not too long ago with the person
> who was having similar problems, but the suggestions didn't seem to
> apply (I use Tween 20 in my PBS rinses, and we use APEX slides from
> Surgipath-not Fischer Plus). I've tried everything I can think of and
> I hope someone may have some other ideas. For reference this is for a
> veterinary diagnostic lab, not a human lab, and this is the only stain
> we've been having this issue with.
>
> My protocol is as follows: After deparfinization/rehydration, AR for
> 20 min in the steamer in Biogenex Citra Solution (I've gotten new
> Citra Solution, tried a pressure cooker)
> 5 min peroxide quench, followed by a 10 min protein block using Dako's
> Protein Block-Serum Free 60 min Primary incubation (Dako CD79a) at 37
> C (decreased dilution from 1:100 to 1:65, new bottle of antibody) 30
> min incubation at room temp of ImmPRESS anti-Mouse Reagent from Vector
> Develop stain with DAB chromogen from Vector
>
> Washes of PBS/Tween 20 follow between steps
>
> Thank you,
> Jennifer Hill
> Research Assistant
> Kansas State University
> Veterinary Diagnostic Lab
>
>
>
> ------------------------------
>
> Message: 7
> Date: Fri, 29 Oct 2010 17:34:50 -0500
> From: Laura.Miller <@t> leica-microsystems.com
> Subject: [Histonet] Laura Miller is Out of the Office.
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID:
>
> <OFDDC515A4.934C900D-ON862577CB.007C0A48-862577CB.007C0A48 <@t> leica-micro
> systems.com
> >
>
> Content-Type: text/plain; charset=US-ASCII
>
>
> I will be out of the office starting 10/29/2010 and will not return
> until 11/01/2010.
>
> I am out of the office for the rest of the day. I will be back on
> Monday!
>
>
> ______________________________________________________________________
> This email has been scanned by the MessageLabs Email Security System.
> For more information please visit http://www.messagelabs.com/email
> ______________________________________________________________________
>
>
>
> ------------------------------
>
> Message: 8
> Date: Fri, 29 Oct 2010 16:38:02 -0600
> From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
> Subject: RE: [Histonet] RE: breast fixation times
> To: "'Kuhnla, Melissa'" <Melissa.Kuhnla <@t> chsli.org>, "'Feher,
> Stephen'"
> <sfeher <@t> CMC-NH.ORG>, "'Tench, Bill'" <Bill.Tench <@t> pph.org>,
> "'Weems,
> Joyce'" <JWeems <@t> sjha.org>, <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <0AD2F85778424E90B81B4CA2D469CDDC <@t> prueggihctechlt>
> Content-Type: text/plain; charset="us-ascii"
>
> Melissa,
>
> Just because the clone you are using is not recommended in the
> guidelines does not mean that you cannot use it. You have to validate
> it the same as using a clone they do recommend, but the bottom line is
> you can use what ever you want in what ever way you want as long as
> you validate the protocol on samples generated in your lab. If you
> want to use a fixation or processing different from what is
> recommended you have to compare it to formalin fixation and "standard"
> (whatever that is) paraffin processing, you would have to do a side by
> side comparison on the same tissue. As long as you follow the
> fixation and processing guidelines you can use the ab u want to use
> without comparing it to abs they recommend, as far as I can tell.
> You must validate that ab on at least 25 samples generated in your
> institution just as you would have to validate an antibody they do
> recommend.
>
> Regards,
>
> Patsy
>
>
>
> Patsy Ruegg, HT(ASCP)QIHC
> IHCtech
> 12635 Montview Blvd. Ste.215
> Aurora, CO 80045
> 720-859-4060
> fax 720-859-4110
> www.ihctech.net
> www.ihcrg.org
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
> Kuhnla, Melissa
> Sent: Friday, October 29, 2010 3:51 AM
> To: Feher, Stephen; Tench, Bill; Weems, Joyce;
> histonet <@t> lists.utsouthwestern.edu
> Subject: RE: [Histonet] RE: breast fixation times
>
> One more question regarding ER/PR/Her2..who am I kidding, we will be
> talking about them forever!! With regards to the ER/PR publication:
> The
> PR clone (Ventana 1E2) that we currently use is not listed as
> 'acceptable'. Is anyone else in this situation? Are you switching to
> another clone?? Have you come across anything speaking of this
> scenario?
> What to do if you currently run a clone not mentioned? It will be a
> relatively easy validation for me, but I can't image I am alone here.
> Thanks, Melissa
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Feher,
> Stephen
> Sent: Thursday, October 28, 2010 5:50 PM
> To: Tench, Bill; Weems, Joyce; histonet <@t> lists.utsouthwestern.edu
> Subject: RE: [Histonet] RE: breast fixation times
>
> Great discussion, comprehensive yet concise. Thanks Bill and Joyce
> and Melissa.
>
>
> Steve
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Tench,
> Bill
> Sent: Wednesday, October 27, 2010 12:26 PM
> To: Weems, Joyce; histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] RE: breast fixation times
>
> My apologies for not including the updates accurate for ER and PR.
>
>
> Bill Tench
> Associate Dir. Laboratory Services
> Chief, Cytology Services
> Palomar Medical Center
> 555 E. Valley Parkway
> Escondido, California 92025
> Bill.Tench <@t> pph.org
> Voice: 760- 739-3037
> Fax: 760-739-2604
>
> -----Original Message-----
> From: Weems, Joyce [mailto:JWeems <@t> sjha.org]
> Sent: Wednesday, October 27, 2010 9:22 AM
> To: Tench, Bill; histonet <@t> lists.utsouthwestern.edu
> Subject: RE: breast fixation times
>
> Thanks for this good explanation, Bill.
>
> One can not follow the guidelines and document the variant in the
> report, but not following them could hurt the patient if there is a
> clinical trial they might participate in. Clinical trials follow the
> protocol to the letter and if the FDA requirement is not met, the
> patient can not participate.
>
> The times were extended for ER and PR to 72 hours, but NOT yet for
> Her2.
> So...because the tissue is all the same, we must follow the 48 hour
> limit. We just had a case this weekend. Had the clinical staff remove
> it from the processor on Sun morning and embedded it Monday. We don't
> ususally have this problem as we are a 6-day lab, but it was finished
> too late on Fri.
>
> Cheers,j
>
> Joyce Weems
> Pathology Manager
> Saint Joseph's Hospital
> 5665 Peachtree Dunwoody Rd NE
> Atlanta, GA 30342
> 678-843-7376 - Phone
> 678-843-7831 - Fax
>
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Tench,
> Bill
> Sent: Wednesday, October 27, 2010 11:50
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] breast fixation times
>
> There is no exception for core biopsies, as reasonable as that may
> seem.
> I have had that discussion with the purveyors of the guidelines. 6-48
> is the current standard. there was a lot of discussion about
> exceeding
> 48 and using the FISH option. My colleague responsible for this
> wrote:
> It is in the CAP checklist, ANP 22998:
>
> If the laboratory assesses Her2 by IHC or Her2 gene amplification by
> in-situ hybridization (FISH, CISH, SISH), does the lab have a
> documented procedure for ensuring appropriate length of fixation of
> specimens tested?
>
> Specimens subject to Her2 testing should be fixed in 10% neutral
> buffered formalin for at least 6 hours and no longer than 48 hours.
> While fixation outside of these time limits is not an absolute
> exclusion criterion for Her2 testing, labs should qualify any negative
> results for specimens fixed less than 6 hours or longer than 48 hours.
> For cases with negative results by IHC, consideration should be given
> to performing confirmatory analysis by in-situ hybridization.
>
> There is also a table in the original ASCO/CAP Guideline
> Recommendations for Her2 in Breast Cancer (Arch Pathol Lab Med, Vol
> 131, Jan 2007) that states that tissue fixed in formalin for greater
> than
> 48
> hours is not an absolute exclusion criterion, but if known to be fixed
> longer than 48 hours or unknown, the report should qualify any
> negative result with this information (table 6).
>
> As for upcoming changes, i don't know other than these time
> limitations are suppose to be more rigorously applied to ER and PR,
> along with the newly instituted documentation of time between excision
> and time placed in fixative.
>
>
> Bill Tench
> Associate Dir. Laboratory Services
> Chief, Cytology Services
> Palomar Medical Center
> 555 E. Valley Parkway
> Escondido, California 92025
> Bill.Tench <@t> pph.org
> Voice: 760- 739-3037
> Fax: 760-739-2604
>
>
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>
> ------------------------------
>
> Message: 9
> Date: Fri, 29 Oct 2010 22:10:21 -0400
> From: historsd <@t> aol.com
> Subject: [Histonet] cryostat question
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <8CD45EEBD9B9E32-EA4-3DF2 <@t> webmail-d038.sysops.aol.com>
> Content-Type: text/plain; charset="us-ascii"
>
>
> Does anyone have 'hands on' experience performing Mohs using a
> countertop cryostat? Can the specimen head be manipulated to
> acccomodate a badly embedded block? Does the machine stay cold enough
> to perform Mohs? Any advice would be appreciated.
> My e-mail is historsd <@t> aol.com. Thanks, Susan
>
>
> ------------------------------
>
> Message: 10
> Date: Fri, 29 Oct 2010 22:44:58 -0700
> From: Jennifer MacDonald <JMacDonald <@t> mtsac.edu>
> Subject: Re: [Histonet] IHC Staining Problem
> To: Jennifer Hill <jhill <@t> vet.k-state.edu>
> Cc: histonet <histonet <@t> lists.utsouthwestern.edu>,
> histonet-bounces <@t> lists.utsouthwestern.edu
> Message-ID:
>
> <OF83C059C1.879738A5-ON882577CC.001F8665-882577CC.001F9C5A <@t> mtsac.edu
> >
> Content-Type: text/plain; charset="US-ASCII"
>
> How far in advance are your controls being cut? Are your patient
> tissues staining?
>
>
>
>
> Jennifer Hill <jhill <@t> vet.k-state.edu>
> Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
> 10/29/2010 01:11 PM
>
> To
> histonet <histonet <@t> lists.utsouthwestern.edu>
> cc
>
> Subject
> [Histonet] IHC Staining Problem
>
>
>
>
>
>
> I hope someone can help me with a problem I've been having with my
> CD79a hand stain. This is a stain that was validated and working
> perfectly, when a few months ago I started having reduced, to no
> staining on my controls, with the occasional run staining a little
> stronger. I followed the discussion not too long ago with the person
> who was having similar problems, but the suggestions didn't seem to
> apply (I use Tween 20 in my PBS rinses, and we use APEX slides from
> Surgipath-not Fischer Plus). I've tried everything I can think of and
> I hope someone may have some other ideas. For reference this is for a
> veterinary diagnostic lab, not a human lab, and this is the only stain
> we've been having this issue with.
>
> My protocol is as follows: After deparfinization/rehydration, AR for
> 20 min in the steamer in Biogenex Citra Solution (I've gotten new
> Citra Solution, tried a pressure cooker)
> 5 min peroxide quench, followed by a 10 min protein block using Dako's
> Protein Block-Serum Free 60 min Primary incubation (Dako CD79a) at 37
> C (decreased dilution from 1:100 to 1:65, new bottle of antibody) 30
> min incubation at room temp of ImmPRESS anti-Mouse Reagent from Vector
> Develop stain with DAB chromogen from Vector
>
> Washes of PBS/Tween 20 follow between steps
>
> Thank you,
> Jennifer Hill
> Research Assistant
> Kansas State University
> Veterinary Diagnostic Lab
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> End of Histonet Digest, Vol 83, Issue 46
> ****************************************
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